symposium a healthy food chain for a balanced … · green and golden tides decline of kelp ......
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SYMPOSIUM A healthy food chain for a
balanced microbiome, Thanks to algae!
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Seaweed microbiome: Importance for cultivation, food & biotechnology
Philippe POTINResearch director at CNRS / Roscoff Biologic station
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A hot spot of biodiversity for understanding the seaweed microbiome
3
Philippe Potin, G. Michel, M. Czjzek, F. Thomas, R.
Larocque, C. Leblanc, A. Gobet, J. Tapia, S. Dittami,
JH Hehemann, G. Correc, C. Boyen, B. Kloareg
.
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Green and golden tides
Decline of kelp
forests worlwide
Impact on coral
reefs
Alginate extraction/
Biotechnology
The microbiome
functions?
Why understanding the seaweedmicrobiome?
Seaweed Food and Feed
Seaweed Cultivation
4
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As humans and all eukaryotes do, seaweeds interact with microbes
5
Matsuo et al. 2005 Science: Thallusineffect on Ulva pertusa
Kumar et al. 2016
18, 3962–3975
• Bacteria shape seaweed
morphology
• Nitrogen assimilation
• Vitamins
• Defense/diseases…
Multiple
opportunistic
pathogens can
cause a bleaching
disease in the red
seaweed Delisea
pulchra
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The seaweed microbiome: how it looks like?
Bacte
rialb
iofilm
Seaweed
Egan et al. FEMS Microbiology reviews 37(3) ·2012
S. Salaün 2010.
L. digitata biofilm by Scanning
Electron Microscopy
and by Fluorescence Confocal
Microscopy in flow cells.
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Seaweed and associated bacteria in a changingenvironment : a complex system = phycobiome
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CHALLENGES:
• Proofs of the role of bacteria in
adaptation
• Learn how to study a complex
system
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Unraveling the Functions of the Macroalgal Microbiome?
Egan et al. FEMS Microbiology reviews 37(3) ·2012
Bacte
rialb
iofi
lm
Seaweed
Cultivable Surface Microbiota
Uncultivable Surface Microbiota « Omics » science revolution
Singh & Reddy, 2016
Front. Microbiol., | https://doi.org/10.3389/fmicb.2015.01488
8
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Bacteria-induced growth and morphogenesis in the green macroalga genus Ulva
Wichard T
Front. Plant Sci.,
03 March 2015 |
9
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Metabolomics of bacteria-induced growth and morphogenesis in Ulva mutabilis (compressa)
Taghreed Alsufyani,
Anne Weiss
and Thomas Wichard
Mar Drugs 2017
10
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Bacteria-induced growth and morphogenesis in the brown alga Ectocarpus sp.
There is a clear developmental difference between the
axenic and non-axenic Ectocarpus sp.
Non-axenic Axenic
Under sterile
conditions there is
not development of
upright filament in
Ectocarpus axenic
strain
Tapia et al.,
2016
Front.
Microbiol.
11
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Culture-dependent techniqueExperimental strategy to isolate and identify bacteria
associated with Ectocarpus sp.
Grinding
Plating PCR16S rRNA
Sequencing
Sequence analysis
Tapia et al., 2016
Front. Microbiol.
Bacteria-induced growth and morphogenesis in the brown alga Ectocarpus sp.
12
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Isolate ID Closest matching strain in BLAST Class % Sequence similarity
Z8_a Marinobacter sp. γ-Proteobacteria 100
R13 Sulfitobacter sp. α-Proteobacteria 100
Es32869_2 Marinobacter sp. γ-Proteobacteria 99
R15 Sulfitobacter sp. α-Proteobacteria 99
Z3 Halomonas sp. γ-Proteobacteria 99
Es32869_1 Methylophaga sp. γ-Proteobacteria 99
R11_a Antarctobacter heliothermus α-Proteobacteria 99
R4_b Alteromonas sp. γ-Proteobacteria 99
Z1 Alteromonas genovensis γ-Proteobacteria 99
Z9_b Bacterium DG940 γ-Proteobacteria 98
R2_b Halomonas sp. γ-Proteobacteria 98
R6_a Bermanella marisrubri γ-Proteobacteria 97
Es534869_
2 Halomonas sp. γ-Proteobacteria 96
R10_a Roseobacter sp. α-Proteobacteria 96
Z8_b Marinobacter sp. γ-Proteobacteria 90
Bacterial isolates identify by 16rRNA gene sequencing.
Tapia et al., 2016Front. Microbiol.
Bacteria-induced growth and morphogenesis in the brown alga Ectocarpus sp.
13
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Non-axenic Axenic Z8a
Bacterial effect on sporulation
Tapia et al., 2016
Front. Microbiol.
Bacteria-induced growth, morphogenesis and reproduction in the brown alga Ectocarpus sp.
14
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Tapia et al., 2016Front. Microbiol.
Waterborne cues in bacteria-induced growth and morphogenesis in the brown alga Ectocarpus sp.
15
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weeks
Dittami et al. ISME J. 2015
Seaweed in
seawater
Seaweed in
freshwater
Algue traitée
aux
antibiotiques en
eau douce
0 3 5
Controls that exclude direct
effect of antibiotics
86 bacterial taxa
Host–microbe interactions as a driver of acclimation to salinity gradients in Ectocarpus subulatus
16
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An example of complementarity: Biosynthesis of vitamin B5
β-alanine Pantoate
Esi0070_0043
Phect3248
EC 6.3.2.1
Vitamine B5
(Acide pantoïque)
Phosphopantothenate
Esi0031_0150
Phect2770
EC 2.7.1.33
Synthèse de Coenzyme A
Phect502
EC 4.1.1.11
Aspartate 2-oxoisovalerate
Ketopantoate
Esi0133_0041
EC 1.1.1.169
Phosphopantothenate biosynthesis Iβ-alanine biosynthesis III
Phect2640
Esi0112_0042
EC 2.1.2.11
Ectocarpus
Pyruvatibacterectocarpi
Dittami et al. 17
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From: Seaweed–microbial interactions: key functions of seaweed-associated bacteriaFEMS Microbiol Ecol. 2014;88(2):213-230. doi:10.1111/1574-6941.12297
FEMS Microbiol Ecol | © 2014 Federation of European Microbiological Societies
Key functions of the seaweedmicrobiome
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The kelp microbiome: a first metagenomics approach
MetaSort untangles metagenome assembly
by reducing microbial community complexity
Peifeng Ji, Yanming Zhang,
Jinfeng Wang & Fangqing Zhao
Nature Communications 8,:
14306 (2017)
•doi:10.1038/ncomms14306
(a) Phylogenetic tree of 75 assembled
kelp bacterial genomes belonging to five phyla.
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The kelp microbiome is very distinct than the surroundingocean and human gut microbiomes
(b) The enriched EGGNOG
functions of horizontally
transferred genes.
Comparison of the KEGG pathways
Ji et al. (2017)
Nature Communications 8,:
14306
20
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The kelp microbiome is very distinct than the surroundingocean and human gut microbiomes
(c) Comparison of the number of CAZyme genes among
the three microbiomes.
(d) Features of the three kelp bacterial genomes (HD112,
HD029 and HD041) assigned to the Planctomycetaceae
family.
Genome size is shown in curved line, and the number of
CAZyme genes in each genome is shown in bar plot.
Ji et al. (2017)
Nature Communications 8,:
1430621
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From genome toenzymatic tools
crystallization
biochemical characterization
Marinebacteria GH16 CBM UNK
medium throughput expression
3D structure
functional characterization
Our current strategy for the discovery of enzymes that degrade seaweed polysaccharides
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Expression and characterisation of a ulvan lyase from the marine bacterium Persicivirga ulvanivorans
23
Ulvan primary structure
P. ulvanivorans colonies on agar plate; Degradation of a 4% ulvan plate; Activity of recombinant ulvan lyase.
Pi Nyvall Collén et al., J. Biol. Chem., 2011
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Zg3628 AgaF
Zg3640 AgaG
Zg1017 AgaH
Zg3376 AgaI
Zg2600 AgaE
Zg4267 AgaC
Zg4203 AgaA
Zg3573 AgaB
Zg4243 AgaD
Zg236 CgkA
Zg2431 LamA
Zg367 LamB
Zg1010 LamC
Zg1014 LamD
Zg1021
Z3347
LicA Bl
GH16 X70 UNK
GH16
GH16 CBM6
GH16
GH16 UNK
GH16 UNK
GH16
GH16
GH16
GH16 CBM16
GH16PKD
GH16
GH16 UNK CBM42
GH16 CBM6
GH16 CBM6 UNK
GH16
b-agarases
k-carrageenases
laminarinases
heterologous over expression of
Zg1017 & Zg2600
?
24
Phylogenetic analysis of Zobelliaenzymes from family GH16
Hehemann JH, Correc G, Barbeyron T, Helbert W, Czjzek M, Michel G. Nature. 2010
464:908-12.
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Activity screening on algal cell wall extracts
2
5
• correlation with increasing L-galactose-6-sulfate units• maximum activity on red algae Porphyra sp.
Hehemann JH,
Correc G,
Barbeyron T,
Helbert W,
Czjzek M,
Michel G.
Nature. 2010
464:908-12.
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The intestinal bacteria of Japanese people received a complete porphyranolytic system from a marine Bacteroidetes
6 of the genes in B. plebeiusare conserved with marine bacteria only
Hehemann JH, Correc G, Barbeyron T, Helbert W, Czjzek M, Michel G. Nature. 2010
464:908-12.
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A genetic gift for sushi eaters
Sushi
Nori
Porphyra/PyropiaGut-metagenome analysis shows thatporphyranases and agarases:
• are frequent in Japanese gut microbiota
• are absent in American gut microbiota
Dietary seaweed is the most probable vector for contact with marine microbes that led to the HGT
Transfer of carbohydrate-active enzymes from marine bacteria to Japanese
gut microbiota.Hehemann JH, Correc G, Barbeyron T, Helbert W, Czjzek M,
Michel G. Nature. 2010 464:908-12.
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The intestinal bacteria of Japanese and Japanese-Hawaianpeople received a complete alginolytic system from the marine Bacteroidetes Zobellia
Characterization of the first alginolytic operons in a marine bacterium: from their emergence in marine Flavobacteria to their independent transfers to marine Proteobacteria and human gut Bacteroides
Thomas et al 2012 Env
Microbiol
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From the seaweed microbiome towardbiotechnology and biorefineries
- Enzyme/protein engineering
- Enzyme-assisted hemisynthesis
- Controled production of chemicals and bioactive molecules
- Bioprocessing of the biomass using enzymes
Probes, markers
optimisation/thermostability
“ glycosynthases ”
ISIAlgolife29
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Thank youfor your attention!
This project has received
funding from the
European Union’s Horizon
2020 research and
innovation programme
under Grant Agreement
no. 727892 (GENIALG).